Rapid Amplification of cDNA Ends (RACE) is a technique used to obtain the full-length sequence of an RNA transcript. RACE can provide the sequence of an RNA transcript from a small known sequence within the transcript all the way to the 5' end (5' RACE-PCR) or 3' end (3' RACE-PCR) of the RNA.

The SMARTer RACE 5’/3’ Kit is an improved version of the SMARTer RACE cDNA Amplification Kit. The SMARTer RACE 5’/3’ Kit provides improved sensitivity, less background and higher specificity over the previous generation of kits. The first-strand cDNA generated with this kit can be used directly in 5’- and 3’-RACE PCR reactions. The SMARTer protocol does not involve any of the adaptor ligation steps that other RACE kits incorporate, making the protocol shorter and significantly easier to use. The new kit is also designed to accommodate larger RNA input volumes (up to 11 µl) and has been shown to perform more efficiently on challenging targets (e.g., those that are long, GC-rich, etc.). The kit includes our highly robust SeqAmp DNA Polymerase for amplification of products, and the In-Fusion HD Cloning Kit for efficient cloning of RACE fragments in the provided linearized vector. The SMARTer RACE 5’/3’ procedure combines SMART first-strand cDNA synthesis technology with a powerful suppression PCR protocol that vastly reduces the background amplification commonly associated with RACE protocols.

Easy to UseThe SMARTer RACE protocol is performed in a single tube, and minimizes handling of both your RNA sample and your synthesized cDNA. Total hands-on time is only four hours.

Complete KitThe SMARTer RACE 5’/3’ Kit contains all the necessary components (save gene-specific primers) to streamline your process. With this single kit, you can begin first-strand cDNA synthesis and proceed through cloning RACE fragments, recovering successful clones on day two.

Requires Only 10 ng of Total RNAOur SMARTer RACE method allows you to utilize small samples, including biopsies, tissue dissections, needle aspirations, and embryonic and rare disease tissues. This optimized protocol significantly reduces non-specific background, and such a reduction is essential when handling very small sample sizes.

Specific Enrichment for 5' EndsOur carefully designed, chemically-treated SMARTer Oligo preferentially hybridizes to the 5' ends of the cDNA being synthesized. Using this SMARTer Oligo, our procedure enriches cDNA pools for 5' sequences, thus increasing the likelihood you will amplify the entire sequence of your gene or the upstream regulatory regions.

No RNA Pretreatment RequiredOur SMARTer RACE method requires no RNA pretreatment with DNase. This protocol works with total RNA, as well as with samples that may be contaminated with genomic DNA.

Related Products

Marathon cDNA Amplification Kit

The Marathon cDNA Amplification Kit method employs a specially-designed adaptor that significantly reduces background and permits both 5'- and 3'-RACE reactions (1, 2) to be performed using the same template. Marathon cDNA amplification can be used to quickly characterize multiple RNAs identified by expressed sequence tags (ESTs), differential display, RNA fingerprinting, or cDNA subtraction. Marathon cDNA synthesis begins with poly A+ RNA and a modified lock-docking oligo(dT) primer that contains two degenerate nucleotides at the 3' end. These nucleotides position the primer at the beginning of the poly A+ tail, eliminating the 3' heterogeneity inherent with conventional oligo(dT) priming. Following cDNA synthesis, blunt ends are created and the Marathon Adaptor is ligated to both ends of the double-stranded cDNA.

RACE-READY cDNAs

Clontech also offers RACE-Ready cDNAs, which are double stranded cDNAs made from high-quality Premium Poly A+ RNA and ligated to the Marathon Adaptor. These cDNAs are ready for 5'- and 3'-RACE PCR and are available from a wide range of tissues and cell types.

Applications

References

Bertling, W. M., et al. (1993) PCR Methods Appl.3:95–99.

Frohman, M. A. (1991) Methods Enzymol.218:340–362.

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.